In vitro interactions between bone marrow stromal cells and hippocampal slice cultures

Abstract

Bone marrow stromal cells (BMSCs) are capable of differentiating into various cell types including brain cells. Several groups have also demonstrated trophic effects of MSC grafts in experimental ischemia models. However, the underlying molecular mechanisms of these effects are not fully understood. We developed an ''in vitro graft model'' which consisted in a coculture of GFP-expressing BMSCs and hippocampal organotypic slice cultures. Total marrow cells (MCs) or BMSCs after one (BMSC 1P ) or five passages (BMSC 5P ) were transplanted on hippocampal slices. During the 10 days of our experiments, MCs and BMSC 1P migrated toward the tissue, but their total number remained constant. Conversely, the number of BMSC 5P decreased over the 10 days of the experiment, and no migration could be detected. Using immunohistochemistry, we observed that the hippocampal slices induced the expression of neural antigens in very few grafted cells, but MCs and BMSC 1P improved the conservation of the hippocampal slice culture. Similar experiments using BMSC 5P did not produce any significant change. We conclude that the number of passages greatly influence BMSCs survival rate, migration and neuroprotective capacities.

Organotypic cultures as tools for optimizing central nervous system cell therapies

“…Three days post-grafting an increase of the length of the axon-like processes was observed as well as the number of neuronal-positive cells, which increased when retinoic acid was added into the culture medium (Abouelfetouh et al, 2004). In the same way, grafted MSCs could migrate into hippocampal tissue and differentiated into neuronal cells under the microenvironment pressure, but the differentiation rate remained very low (Charriere et al, 2010). As expected, NSC grafts within the striatum integrated morphologically forming part of the 3D cytoarchitecture.…”

“…This 3D environment has tremendous value for evaluating the efficacy of cell therapy, by providing external mechanical inputs, interactions between structures and cell adhesion parameters, which all profoundly affect intracellular signalling. In addition, they enable an easier way to study local cell implantation sites, grafted cell migration and also their interaction with the host environment, mainly glial cells and neurons (Abouelfetouh et al, 2004;Charriere et al, 2010;Jaderstad et al, 2010aJaderstad et al, , 2010bJaderstad et al, , 2011Meng et al, 2011;Sarnowska et al, 2009a;Tanvig et al, 2009;Tonnesen et al, 2011). They are advantageous as they allow patch-clamp studies of neuronal electrophysiology (Finley et al, 2004;Jaderstad et al, 2010b;Kearns et al, 2006;Tonnesen et al, 2011) or studies on the influence of calcium or magnesium on the cells and their correlation with survival or apoptosis markers (Bickler and Fahlman, 2004;Turner et al, 2007).…”

In vitro expansion impaired the stemness of early passage mesenchymal stem cells for treatment of cartilage defects

“…12 These results indicate that expansion might attenuate the stemness of MSCs, thereby contributing to reduced therapeutic potential. It has been verified that monolayer culture greatly influences cell behavior, 13 resulting in cell senescence and impairing multipotency. 14 In serial passage of MSCs, telomere activity and chromosome heteromorphosis increase over time.…”

Effects of infrasound on the growth of bone marrow mesenchymal stem cells: A pilot study

“…The bone marrow was flushed from the shaft with Dulbecco's modified Eagle's medium (DMEM)/F12 (Hyclone, Waltham, MA, USA) using a 20-gauge needle. The bone marrow suspension was disaggregated by pipetting several times and the cells were collected by centrifugation (2,200 x g, 5 min) (17). The cells were then cultured in DMEM/F12 with 10% fetal bovine serum (Gibco-BRL, Carlsbad, CA, USA) containing 100 µl/ml penicillin-streptomycin in 25 cm 2 cell culture flasks (Corning Inc., Acton, MA, USA).…”